TALEN-mediated precise genome modification by homologous recombination in zebrafish

Zu, Yao; Tong, Xiangjun; Wang, Zhanxiang; Liu, Da; Pan, Ruochuan; Li, Zhe; Hu, Yingying; Luo, Zhou; Huang, Peng; Wu, Qian; Zhu, Zuoyan; Zhang, Bo; Lin, Shuo

doi:10.1038/nmeth.2374

Cited by 308 publications

(233 citation statements)

References 19 publications

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“…Gene targeting via non-homologous end-joining (NHEJ) has also been suggested to generate combined reporter and knockout alleles 9 . Combination of TALEN-based gene targeting with insertion of a few base pairs or transgenes using small ssDNA oligonucleotides or donor vectors with longer homologous arms was initially reported for zebrafish 10,11 and later extended to other animal models including mouse 12,13 . The mouse remains the most widely used animal model in the study of human biology and diseases.…”

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confidence: 99%

Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases

et al. 2014

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Generation of mouse models by introducing transgenes using homologous recombination is critical for understanding fundamental biology and pathology of human diseases. Here we investigate whether artificial transcription activator-like effector nucleases (TALENs)-powerful tools that induce DNA double-strand breaks at specific genomic locations-can be combined with a targeting vector to induce homologous recombination for the introduction of a transgene in embryonic stem cells and fertilized murine oocytes. We describe the generation of a conditional mouse model using TALENs, which introduce double-strand breaks at the genomic locus of the special AT-rich sequence-binding protein-1 in combination with a large 14.4 kb targeting template vector. We report successful germline transmission of this allele and demonstrate its recombination in primary cells in the presence of Cre-recombinase. These results suggest that TALEN-assisted induction of DNA double-strand breaks can facilitate homologous recombination of complex targeting constructs directly in oocytes.

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confidence: 99%

Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases

et al. 2014

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“…Novel approaches have been recently designed to efficiently produce animals that are genetically altered in specific genomic sequences; zinc-finger nuclease (ZFN) (8)(9)(10) and transcription activator-like effector nuclease (TALEN) (11)(12)(13)(14)(15)(16) are representative next-generation platforms for customized genomic editing in transgenic animals, as well as in cultured cells in vitro. ZFNs and TALENs containing a DNA-binding domain and Fok I nuclease domain are currently crucial tools to dissect individual gene function in complicated biological systems with targeted genetic deletions and alterations.…”

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confidence: 99%

Targeted gene knockout in chickens mediated by TALENs

Park

Lee

Kim

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

151

138

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Genetically modified animals are used for industrial applications as well as scientific research, and studies on these animals contribute to a better understanding of biological mechanisms. Gene targeting techniques have been developed to edit specific gene loci in the genome, but the conventional strategy of homologous recombination with a gene-targeted vector has low efficiency and many technical complications. Here, we generated specific gene knockout chickens through the use of transcription activator-like effector nuclease (TALEN)-mediated gene targeting. In this study, we accomplished targeted knockout of the ovalbumin (OV) gene in the chicken primordial germ cells, and OV gene mutant offspring were generated through test-cross analysis. TALENs successfully induced nucleotide deletion mutations of ORF shifts, resulting in loss of chicken OV gene function. Our results demonstrate that the TALEN technique used in the chicken primordial germ cell line is a powerful strategy to create specific genome-edited chickens safely for practical applications.genetic modification | programmable genome editing | germ-line chimera | poultry | egg protein

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“…More recently, the transcription activator-like effectors (TALEs) technology, a milestone in the development of zebrafish mutant and transgenic lines, has lifted the limit of loci-specific targeting. With very low off-targeting effects, TALEs were therefore the first successful genome editing method that permitted homologous-directed recombination (HDR) and NHEJ-mediated knockin in zebrafish (Bedell et al 2012;Zu et al 2013). Two reports (Chang et al 2013;Hwang et al 2013b) showed that double stranded breaks (DSB), which are simpler in design and have higher mutagenesis efficiency, could also be generated using the Crispr/Cas9 technology based on the same approach used by Bedell et al (2012).…”

Section: Crispr/cas9-mediated Knockin Approaches In Zebrafishmentioning

confidence: 99%

CRISPR/Cas9-Mediated Knockin and Knockout in Zebrafish

Albadri

Santis

Donato

et al. 2017

Research and Perspectives in Neurosciences

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The zebrafish (Danio rerio) has emerged in recent years as a powerful vertebrate model to study neuronal circuit development and function, thanks to its relatively small size, rapid external development and translucency. These features allow the easy application of in vivo microscopy analysis and optical perturbation of neuronal function. So far, genetic manipulation in zebrafish has been limited to the generation of constitutive loss-of-function alleles and transgenic models.

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TALEN-mediated precise genome modification by homologous recombination in zebrafish

Cited by 308 publications

References 19 publications

Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases

Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases

Targeted gene knockout in chickens mediated by TALENs

CRISPR/Cas9-Mediated Knockin and Knockout in Zebrafish

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